Three‐Dimensional Mid‐Infrared Tomographic Imaging of Endogenous and Exogenous Molecules in a Single Intact Cell with Subcellular Resolution. Issue 1 (13th November 2014) - Quaroni, Luca
Abstract: Microscopy in the mid‐infrared spectral range provides detailed chemical information on a sample at moderate spatial resolution and is being used increasingly in the characterization of biological entities as challenging as single cells. However, a conventional cellular 2D imaging measurement is limited in its ability to associate specific compositional information to subcellular structures because of the interference from the complex topography of the sample. Herein we provide a method and protocols that overcome this challenge in which tilt‐series infrared tomography is used with a standard benchtop infrared microscope. This approach gives access to the quantitative 3D distribution of molecular components based on the intrinsic contrast provided by the sample. We demonstrate the method by quantifying the distribution of an exogenous metal carbonyl complex throughout the cell and by reporting changes in its coordination sphere in different locations in the cell. Abstract : Now in 3D : Conventional 2D cell imaging methods are limited in their ability to associate specific information on the sample composition with subcellular structures. A new method, in which tilt‐series infrared tomography is used with a standard benchtop infrared microscopy, provides quantitative information on the 3D distribution of molecular components based on the intrinsic contrast provided by the sample.